Figure 4.

Degradation pathways for BEST1 clearance in BD, ARB, and ADVIRC hiPSC-RPEs. Representative Western blot images after α-BEST1 immunoblot staining of whole cell lysates from untreated and treated hiPSC-RPEs of (A) control +/+ #1 and patients associated with BD_IN and ARB, respectively. (B) Summary of BEST1 protein expression obtained from experiments shown in (A). Representative Western blot images after α-BEST1 immunoblot staining of whole cell lysates from untreated and treated hiPSC-RPEs of (C) control +/+ #2 and patients associated with BD_PM and ADVIRC, respectively. (D) Summary of BEST1 protein expression obtained from experiments shown in (C) relative to untreated samples and normalized against β-actin from the same blot. Samples in (A) and (C) were assayed after 12 h of CHX treatment (20 µg/mL) in the absence or presence of proteasomal inhibitor MG132 (40 µM) or lysosomal inhibitor chloroquine (CQ) (100 µM). In (B) and (D) graphs for BD_IN, BD_PM, ARB, and ADVIRC samples are averaged and mean values are given as mean ± SD from two technical replicates per individual sample (n = 13). A summary of hiPSC clones used in the experiments is given in Supplementary Table S4. Statistical analysis was performed applying Kruskal-Wallis test, following post-hoc Dunn’s multiple comparisons test including Benjamini-Hochberg procedure: * = p < 0.05; *** = p < 0.001; n.s. = not significant.